1. Field of Invention
This invention relates to the synthesis of derivative compounds of N-6-trimethyl-L-lysine.
2. Description of Related Art
All material referenced in the prior provisional and non-provisional applications are hereby incorporated by reference.
In the parent application Ser. No. 11/105,165, allowed on Nov. 17, 2010 to issue as a patent, it was shown by carefully compiled experimental results and other scientific demonstration, that therapeutic use of TML may successfully arrest, and in certain respects, reverse the degeneration associated with the group of progressive neurological diseases called neuronal ceroid lipofuscinoses (NCL).
The parent application described the symptoms and common as well as distinct characteristics of the spectrum of NCL group, including the following: Batten Disease, Santavuori disease, Late-Infantile Neuronal Ceroid Lipofuscinoses (LINCL), Speilmeyer-Sjogren disease, Kuf disease, Parry disease, Bernheimer-Seitelberger syndrome, Bielschowsky amaurotic idiocy, Bielschowsky disease, Jansky-Bielschowsky disease, Seitelberger disease, late infantile amaurotic idiocy, late infantile Batten disease, subacute late infantile neuronal ceroid-lipofuscinosis, Zeman-Dyken-Lake-Santavuori-Savukoski disease.
At the genetic level, the neuronal ceroid lipofuscinoses (NCL's) result from mutations in at least eight genes, and these mutations are responsible for causing the various expressions of the neurodegenerative diseases collectively identified as NCLs. A summary background of these mutations and a survey of the background reference literature were given in the parent application. See Table A by Gene Locus.
TABLE ANeuronal Ceroid Lipofuscinosis - Summary of SymptomsSYMPTOMSCLN1CLN2CLN3CLN4CLN5CLN6CLN7CLN8CLN9CLN10DementiaYesYesYesYesYesYesSeizuresYesYesYesYesYesYesYesYesYesyes(hyperkneticmovements,hand/feettremors)Progressive VisualYesYesYesYesYesYesYesNoYesnewbornFailureinfantMental RetardationYesYesYesYesYesYesYesYesLoss Of SpeechYesYesYesYesYesYesyesRegression ofYesYesYesYesYesyesMotorDevelopmentAtaxiaYesYesYesYesYesYesyesMuscularYesYesYesYesyesHypotonia/DystoniaMicrocephalyYesOptic Atrophy/YesYesYesYesMacularDegenerationRetinitisPigmentosaMyoclonusYesYesYesYesYesYesNoCerebellar AtrophyYesYesYesYesyesQuadraparesisYesRefractory EpilepsyYesBehavioralYesYesInvolvement(Anger Outburst,Physical Violence)Table A Notes:1. According to Mole et al..2005, the clinical course of the NCL's include progressive dementia, seizures, and progressive visual failure (Full text available at http://www.springerlink.com/content/xu2406100j81034w/fulltext.pdf ).2. A ‘Yes’ means that the symptom is a characteristic of the disease. A ‘No’ means that the OMIM synopsis from clearly stated that the specific symptom is NOT characteristic of that particular NCL. An empty space for a particular symptom does not necessarily preclude it from being part of the characteristics of that particular NCL; it was not mentioned specifically in the OMIM synopsis. For instance, CLN6 is an LINCL (CLN2) variant. It did not specifically mention Mental Retardation or Loss of Speech or Cerebellar Atrophy; but it would be reasonable that Mental Retardation/Loss of Speech/Cerebellar would be part of the continuum.CLN1 (Infantile)CLN2 (Late Infantile)CLN3 (Juvenile)CLN4a (Kufs Disease)CLN5 (Late Infantile, Finnish Variant)CLN6 (Late Infantile, Variant, Included, Variable age at onset)CLN7CLN8CLN8 (Northern Epilepsy Variant)CLN9CLN10 Cathepsin D-Deficient, Congenital
No generally effective treatment for many of the diseases mentioned above is currently available and these diseases are generally fatal. The line of treatment proposed in the parent application is based on administering, in the form of a therapeutic agent of very high purity, any of the following to a human being in need thereof: (a) N-6-trimethyl-L-lysine, (b) a prodrug thereof, or (c) a pharmaceutically acceptable salt of N-6-trimethyl-L-lysine or said prodrug.
The one common characteristic in all these disorders has been found to be the accumulation of autofluorescent storage material in all tissues, particularly pronounced in the central nervous system. This characteristic has been tied to the fundamental role of L-carnitine in metabolism, such as the prevention of hyperammonimia, lipid peroxidation and fatty acid metabolism. It has also been found that availability of TML, as the rate limiting step in the regulation of feedback inhibition for L-carnitine biosynthesis, is crucial to the biosynthesis of L-carnitine. (Schematic 1, by F. M. Vaz and R. J. A. Wandars, Biochem. J., 361, 417-429, 2002).

A summary of the role of carnitine, which was described in greater detail in the parent application, is outlined below:
(a) From a biochemical standpoint, L-carnitine plays an essential role in energy metabolism. In fatty acid metabolism, it serves as shuttle between the mitochondrial membrane and the mitochondria inner-workings permitting breakdown of the long carbon fragment. A major part of that role is in maintaining a balance between the concentration of a compound called acyl CoA in the cell compartments and in sugar metabolism.
(b) Optimal ATP production in humans from either dietary or stored fatty acids is dependent on L-carnitine. L-Carnitine has several roles, most of which involve conjugation of acyl residues to the b-hydroxyl group of the L-carnitine with subsequent translocation of this complex from one cellular compartment to another.
(c) Defects in fatty acid oxidation are a source of major morbidity and are potentially rapidly fatal. Fatty acid oxidation defects encompass a spectrum of clinical disorders, including recurrent hypoglycemic, hypoketotic encephalopathy or Reye-like syndrome in infancy with secondary seizures and potential developmental delay, progressive lipid storage myopathy, recurrent myoglobinuria, neuropathy, and progressive cardiomyopathy.
(d) Administration of L-carnitine prevents acute ammonia toxicity and enhances the efficacy of ammonia elimination as urea and glutamine. In addition the cytotoxic effects of ammonia, possibly arising from lipid peroxidation, are ameliorated by L-carnitine. These data indicate the feasibility of utilization of L-carnitine in the therapy of human hyperammonemic syndromes.
(e) L-Carnitine deficiency can be defined as a decrease of intracellular L-carnitine, and is a factor, inter alia, in the inhibition of the mitochondrial oxidation of long-chain fatty acids during fasting, heart or liver failure (which may in turn cause encephalopathy by hypoketonemia, hypoglycemia and hyper-ammonium), and lower acetylcholine synthesis in the nervous system.
L-carnitine plays a key, and critical, role in enhancing fat metabolism. It appears evident that L-carnitine works by transporting fatty acids to be burned for fuel, increasing both energy supply and lean muscle mass. Most reports found that unless an individual is deficient in L-carnitine, it is an unnecessary ergogenic aid. This contrasts with an apparent need in case of L-carnitine deficiency (e.g., in the case pursued by the inventors of Late Infantile Neuronal Ceroid Lipofuscinosis—one form of Batten Disease), of the correct operation of the endogenous production of L-carnitine. This need was corroborated in the observations of dogs with Batten Disease given exogenous L-Carnitine (Siakotos A. N., Hutchins G. D., Farlow M. R., Katz M. L., European Journal of Pediatric Neurology 5 Suppl A: 151-6, 2001) and those of the parents of the child who was afflicted with LINCL (discussed below). The child was given exogenous L-carnitine for over three years without significant metabolic changes or marked outward observations of her condition. It was only the delivery of exogenous TML to the afflicted LINCL child that yielded significant metabolic and outward, observable changes to her condition.
A discussion of the experimental results presented in the parent application showed effectiveness of TML therapy in the amelioration of several symptoms including: hyperammonemia, glutamine levels, insomnia, “nervousness” or myoclonus.
L-Carnitine may be essential or “conditionally” essential for several groups of people including: normal infants, premature infants, and both children and adults suffering from a variety of genetic, infectious, and injury-related illnesses. For example, some cardiomyopathies which afflict children are due to metabolic errors or deficiencies. There is data that supports treatment of some myocardial dysfunctions with L-carnitine supplementation. (Winter, S., Joe, K., Prochazka J., Francis, P., Hamilton, W., Linn, L., Helton, E. (1995) J. Child Neurol. 10, Supple 2: S45-51.)
For these and other reasons, all of which were described in detail in the parent application, it is believed that TML, or its derivatives which had been proposed in the parent application, may be used for the treatment of a human being diagnosed with one or more of the following: defects in carnitine biosynthesis pathway, inefficiency of endogeneous processes involving TML, over-accumulation of TML bound protein at the cellular level, renal failure conditions, hyperammonemic encephalopathy, over-accumulation of glutamine in the brain, reduced and deficient fatty acid metabolism and shuttling of fatty acid in to mitochondria, insufficient ATP production or subsequent energy production and all the cellular activities associated with these events, defective fatty acid oxidation resulting from carnitine deficiency, hypoglycemia, hypoketotic, encephalopathy, Reye-like syndrome, for recurrent seizures and developmental delay, AIDS or AIDS-like conditions, over-accumulation of lipids causing myopathy, myoglobinuria, neuropathy, cardiomyopathy, ammonia over-production, hyperammonemic syndromes, over accumulation of triacylglycrols, Batten diseases, infantile neuronal lipofuscinoses diseases (Santavvori diseases), Late infantile neuronal lipofuscinoses diseases (Jansky-Bielscowsky), Speilmeyer disease, Sjorgsen disease, Kuf diseases, Parry diseases, Juvenile or adult neuronal lipofuscinoses diseases (“NCL”) disease, lysosomal accumulation of mitochondrial ATP synthase subunit and associated byproducts, ataxia and seizures, various stages of mental impairment, (e.g., learning disability, clumsiness, stumbling, impaired motor skills, and dementia, hyperandrogenism caused by NCL, defective dopamine receptors caused by NCL, epileptic fits, myoclonic epilepsy, Parkinson's disease, and Alzheimer's disease.
The experimental results presented to prosecute the parent application showed that the progress observed by the medical care-givers to the LINCL child-patient correlated with the administration of high purity TML. See Table B below.
TABLE BResults After TML Therapy.Test NameNov. 19, 2003Clinical RangeJan. 29, 2004Clinical RangeHgb14.4high14normalHCT42high40.3normalRDW11.5high12.3normalABS Lymphocytes2.2low2.5normalGlycine50high25normalTaurine24high19normalCarnitine, Total40normal43normalCarnitine, Esthers7normal10normalAlanine87high47normalCarbon Dioxide32high22normalBUN2low5(low) (6 is norm!)AST60high 50 (high)(40 is norm)Platelets586high461 (high)(369 norm)Glutamine99high70normalNotes to the Table B:(a)HCB = hemoglobin, HCT = Hematocrit, RDW Red Cell Distribution Width, ABS absolute, BUN Blood Urea Nitrogen, AST = Aspartate Amonotransferase)(b)The examining physicians comments of Nov. 19, 2003 regarding Table B: (I) Alanine is elevated, this may be seen in states with increased pyruvate, (ii) Glutamine is increased, this may be seen, with Hyperammonemia.; Clinical correlation is indicated.(c)The examining physicians comments on Jan. 29, 2004 that no significant elevation of serum amino acid was seen.(d)The patient's glucose and potassium increased (Glucose 93 baseline to 132; Potassium 4.4 baseline to 4.8). Even though the follow up blood work was done after an all night fast, we did give her some “Gatorade” to drink before the blood test. This was given with her Klonopin to wash it down and certainly could be a contributing factor to the rise in glucose and potassium.
It was also found during the administration of TML to the child that it was essential that high purity, therapeutic-grade TML be used for treatment purposes.
Therefore, methods of purifying the TML based compounds were invented by the team of current inventors. These inventions are listed below in the present application.